The following reaction mixture was employed: 0.02 m
sodium phosphate buffer,
20 mm catechol, 400 lL of
enzyme extract (pH 6.8); total volume 3.0 mL.
The
mixture was incubated at 30 C and the enzymatic
activity evaluated by measuring the increase in optical
density (OD) at 410 nm (Serradell et al., 2000).